Supplementary data for: Comparison of transcriptomic profiles between HFPO-DA and prototypical PPARa, PPARg, and cytotoxic agents in wild-type and PPARa knockout mouse hepatocytes

Recent in vitro transcriptomic analyses for the short-chain polyfluoroalkyl substance (PFAS), HFPO-DA (ammonium, 2,3,3,3-tetrafluoro-2-(heptafluoropropoxy)-propanoate), support conclusions from in vivo data that HFPO-DA-mediated liver effects in mice are part of the early key events of the peroxisome proliferator-activated receptor alpha (PPARa)activator-induced rodent hepatocarcinogenesis mode of action (MOA). Transcriptomic responses in HFPO-DA-treated rodent hepatocytes have high concordance with those treated with a PPARa agonist and lack concordance with those treated with PPARg agonists or cytotoxic agents. To elucidate whether HFPO-DA-mediated transcriptomic responses in mouse liver are PPARa-dependent, additional transcriptomic analyses were conducted on samples from primary PPARaknockout (KO) and wild-type (WT) mouse hepatocytes exposed for 12, 24 or 72 hours with various concentrations of HFPO-DA, or well-established agonists of PPARa (GW7647) and PPARg (rosiglitazone), or cyt..., Primary Hepatocyte Isolation and Culture
Mouse hepatocytes were isolated from the livers of 11 week-old male B6129SF2/J mice (stock #101045) and 11 week-old male PPARa-null mice (B6;129S4-Pparatm1Gonz/J, stock #008154) purchased from The Jackson Laboratory (Bar Harbor, ME). As described in Lee et al. (1995), PPARa-null mice were generated by a targeted disruption of the ligand-binding domain (i.e., deletion of 83 base pairs in exon 8, see Lee et al. 1995 for details) of the mouse PPARa (mPPARa) gene, rendering the mPPARa gene nonfunctional. Although nonfunctional, mPPARa RNA was detected in these mice at very low expression levels; however, mice completely lack expression of mPPARa protein as measured by Western blotting, and lack functional protein activity as indicated by the inability to activate downstream PPARa target genes (Lee et al. 1995). PPARa-null mice are considered constitutive knockout mice (i.e., PPARa is nonfunctional in the entire animal), thus primary hepatocytes from ..., , # Supplementary data for: **Comparison of transcriptomic profiles between HFPO-DA and prototypical PPARa, PPARg, and cytotoxic agents in wild-type and PPARa knockout mouse hepatocytes**

[https://doi.org/10.5061/dryad.pc866t1wp](https://doi.org/10.5061/dryad.pc866t1wp)

To elucidate whether HFPO-DA-mediated transcriptomic responses in mouse liver are PPARa-dependent, transcriptomic analyses were conducted on samples from primary PPARa knockout (KO) and wild-type (WT) mouse hepatocytes exposed for 12, 24 or 72 hours with various concentrations of HFPO-DA, or established agonists of PPARa (GW7647) and PPARg (rosiglitazone), or cytotoxic agents (acetaminophen or d-galactosamine).

## **List of files**

A list of each file included in this dataset:

·       **Supplementary Files S1 & S2_Sequencing Quality & Cytotoxicity Results.xlsx** – Summary of sequencing quality data and observations of cytotoxicity.

·       **Supplementary File S3a_DEPs_Mouse_B6129SF2J.xlsx** – Differential p...